Immunocytochemical localization of the subunits of glycoprotein hormones (LH, FSH, and TSH) in the bullfrog pituitary gland using monoclonal antibodies and polyclonal antiserum

General and Comparative Endocrinology
S TanakaK Kurosumi

Abstract

Using beta and alpha subunits of bullfrog follitropin (FSH) III (pI 6.2), which were highly purified by HPLC, we generated three monoclonal antibodies (MCAs) to FSH beta subunit (FSH beta) and six to FSH alpha subunit (FSH alpha). They were produced by hybridomas derived from the myeloma X63.Ag8.653 and spleen lymphocytes from mice immunized with each subunit. Non-competitive binding tests revealed that one of the MCAs against FSH beta (BF3B25) bound strongly to intact FSH and its beta subunit, but not FSH alpha, lutropin (LH), LH alpha, and LH beta. The immunoblotting results also showed a similar immunological specificity for BF3B25. Cross-reactivity of bullfrog FSH against BF3B25 was 19.4%, when compared with FSH beta in the competitive inhibition assay system. On the other hand, noncompetitive binding tests and immunoblotting results showed that one of the MCAs against FSH alpha (BF3A20) bound strongly to intact LH and FSH and their alpha subunits, but not their beta subunits. The inhibition curves obtained using the alpha subunits of LH and FSH were similar. In the sexually mature bullfrog pituitary, immunoreactive FSH cells stained with MCA BF3B25 were distributed throughout the pars distalis, except for the rostral regio...Continue Reading

Citations

Oct 20, 1999·General and Comparative Endocrinology·M FiorentinoR K Rastogi
Apr 28, 2011·General and Comparative Endocrinology·Mari SudaMasahisa Nakamura
Jul 1, 1997·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·S TanakaK Wakabayashi
Jul 17, 2012·Sexual Development : Genetics, Molecular Biology, Evolution, Endocrinology, Embryology, and Pathology of Sex Determination and Differentiation·M Nakamura
Feb 21, 2008·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Verónica Filippa, Fabian Mohamed

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